Difference between revisions of "Part:BBa K1022103"
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<partinfo>BBa_K1022103 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1022103 SequenceAndFeatures</partinfo> | ||
− | + | ==Characterization== | |
'''For more info, visit [http://2013.igem.org/Team:TU-Delft/Peptides#SUMO TU Delft iGEM13 Wiki]''' | '''For more info, visit [http://2013.igem.org/Team:TU-Delft/Peptides#SUMO TU Delft iGEM13 Wiki]''' | ||
− | + | ===Peptide production=== | |
'''Introduction:''' | '''Introduction:''' | ||
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− | + | ===MIC determination=== | |
The MIC of peptide on ''S. delphini, B. subtilis and E. coli'' was also done. This characterization can be seen in construct [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1022112 BBa_K1022112]. | The MIC of peptide on ''S. delphini, B. subtilis and E. coli'' was also done. This characterization can be seen in construct [https://parts.igem.org/wiki/index.php?title=Part:BBa_K1022112 BBa_K1022112]. |
Revision as of 13:56, 3 October 2013
pT7: RBS: His6- SUMO: Magainin II
This part codes for the peptide 'Magainin II' tagged with 'His-6-SUMO' molecule. Its production is triggered by IPTG (through pT7 promoter).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 46
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 170
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 355
- 1000COMPATIBLE WITH RFC[1000]
Characterization
For more info, visit [http://2013.igem.org/Team:TU-Delft/Peptides#SUMO TU Delft iGEM13 Wiki]
Peptide production
Introduction:
In the project by iGEM 13 Team TU Delft, a SUMO-peptide fusion was opted as a suitable expression system as they make the fusion proteins more soluble. The peptide by itself is not soluble in the cytoplasm but making a fusion of peptide with Small Ubiquitin like Modifiers (SUMO) will increase the solubility of the peptide, thus increasing the cytoplasmic fraction of the peptide.
A gene was constructed in such a way that the SUMO-peptide production was driven by the strong T7 phage promoter. This gene containing plasmid was harboured in a BL21(DE3) strain that has lac promoter driven T7 polymerase. Upon induction by IPTG the SUMO peptide fusion is produced as a soluble protein fraction.
The protocol can be seen [http://2013.igem.org/Team:TU-Delft/Protocol_10#protocol_10 here].
Result:
Discussion:
MIC determination
The MIC of peptide on S. delphini, B. subtilis and E. coli was also done. This characterization can be seen in construct BBa_K1022112.
Also visit the [http://2013.igem.org/Team:TU-Delft/PeptideCharacterization Peptide characterization] page on TU Delft iGEM13 Wiki to know more about the MIC experiments.